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1. I DANISH MeO TECHNO LG D Auto Dual Wire Myograph System Model 510A User Manual Version 3 3 DANISH OM CT EC kl CC DMT Auto dual wire myograph system Model 510A Auto Dual Wire Myograph System Model 510A User manual Trademarks PowerLab and LabChart are registered trademarks of ADInstruments Pty Ltd The names of specific recording units such as PowerLab 4 25 are trademarks of ADInstruments Pty Ltd Pentium is a registered trademark of the Intel Corporation Windows Windows 95 Windows 98 Windows ME Windows NT Windows 2000 Windows XP and Vista are registered trademarks of Microsoft Corporation All other trademarks are the properties of their respective owners DMT reserves the right to alter specifications as required This document was as far as possible accurate at the time of printing Changes may have been made to the software and hardware it describes since then New information may be supplied separately This documentation is provided with the DMT Auto dual wire myograph system Model 510A v 3 3 Document Number 510A UG3 3A No part of this document may be reproduced by any means without the prior written permission of DMT Copyright 2008 DMT A S M DANISH MYO TECHNOL OG DMT A S DMT Asia DMT USA Inc Skejbyparken 152 Everwin Gardens 1201 Peachtree Street DK 8200 Aarhus N Rm 502 Block B 400 Colony Square Suite 200 630 Denmark 521 Wanping Nan Lu Atlanta G
2. Auto dual wire myograph system Model 510A Figure 4 5 Illustration of how to replace the force transducer Important Notice how the plug is connected to the force transducer so no mistake is made when connecting the plug to the new force transducer 4 5 Carefully disconnect the plug on the force transducer Loosen and remove the two Allen screws B and washers as illustrated in fig 4 5 and carefully remove the old transducer Remove any remaining grease from the transducer pin left inside of the trans ducer compartment of the myograph unit Clean the hole leading from the trans ducer compartment to the myograph chamber Gently place the new force transducer into position Place the Allen screws and washers in their positions ready to be tightened Before tightening the Allen screws be sure that the transducer pin is placed directly in the center of the chamber pinhole when viewing from the side of the myograph chamber Tighten the two Allen screws and replace the bottom plate Tighten the four screws on the bottom plate User manual 510A Chapter 4 33 34 4 4 Myograph maintenance The Auto dual wire myograph 510A system is a very delicate and sophisticated piece of research equipment DMT recommend that the following sections are read care fully and that the instructions are followed at all times 4 4 1 Myograph chamber pipes To prevent the pipes from being blocked by buffer salt deposits after an exper
3. 23 LI OM OB of lt lt 3 TeuoftoM i gt tutofiorm In calculating the ACh in the myograph chamber the applied volume of ACh is ignored 5 8 Buffer Recipes Physiological Saline Solution PSS To make 1 L of PSS Solution 1 Chemical MW g mol NaCl 58 44 KCI 14 56 MgSO 7H 0 246 48 H PO 136 09 Solution 2 Chemical MW g mol CaCl 2H 0 147 02 Solution 3 Chemical MW g mol NaHCO 84 01 EDTA 372 24 Glucose 198 77 Conc mmol L 118 99 4 69 1 17 1 18 Conc mmol L 2 50 Conc mmol L 25 00 0 03 5 50 50 DMT Auto dual wire myograph system Model 510A Conc g L 6 95 0 35 0 29 0 16 Conc g L 0 37 Conc g L 2 10 0 01 1 09 ZS Dissolve the chemicals in approximately 100 mL double distilled H O as three individual solutions as described in the table above Gently heat solution 3 to dissolve the EDTA Solution 1 is added to a graduated bottle and the bottle is filled with double distilled H O to a final volume of 500 mL Solution 3 is added to the graduated bottle which afterwards is filled with additional double distilled H O to a final volume of about 850 mL Aerate the solution with carbogen 95 O 5 CO for about 20 minutes Solution 2 is added and the graduated bottle is filled with additional double distilled H O to reach the final volume of 1000 mL Continue the carbogen bubbling until the pH of the buffer solution reaches 7 4 User ma
4. 3 1 Simple transducer check If the force reading on the Myo Interface appears unstable then first check that the Myo Interface and the Myograph unit are properly connected through the 25 pin grey cable If the force reading still appears unstable then perform a new weight calibration of the force transducer as described earlier in this Chapter During the weight calibration monitor the relative force reading values in the calibra tion menu on the Myo Interface e If the value is O or above 6500 the force transducer is broken and needs to be changed e If the reading is between 1 499 or 3001 6250 then contact DMT for further instructions If the message OFF is displayed in either of the force reading lines in the Main Menu on the Myo Interface then the force transducer is broken and needs to be replaced In case of any other problems related to the force transducer please contact DMT for further instruction and advice 4 3 2 Force transducer replacement In case that the force transducer is broken and needs to be changed please follow this step by step replacement procedure carefully 1 Disconnect the myograph unit from the Myo Interface grey cable 2 Carefully remove the support connected to the transducer pin by loosening the screw on top of the support as illustrated in fig 4 1 1D 3 Turm the myograph unit up side down and remove the bottom plate by loosening the four screws A as illustrated in fig 4 5 1 DMT
5. Press F3 to open the Zero force menu Press F1 to zero NEAR force reading Press F2 to zero FAR force reading Press F3 to zero both NEAR and FAR force readings at the same time Press F4 for the display to show a condensed Main Menu showing NEAR FAR force read ings actual set temperature heat status on off and pH reading optional Mounting menu The mounting menu displays Near pos Far pos Release Position of the NEAR myograph jaws Position of the FAR myograph jaws The readings in um are the actual distance of separation between the jaws The reading in um shows the release set value The setting is the apart distance that the jaws will move on to when commanded to do so in the F2 Release menu as described below Info Change menu Valve on Zero force Condensed menu F2 Activate Valve Yes F1 No F4 F3 F1 Zero FAR force F2 Zero FAR force F3 Zero both F4 F4 Near 12 5mN Far 2 7 MN Act 26 5 Set 37 0 Heat OFF Mounting User manual 510A Chapter 3 19 20 The position of the jaws may also be adjusted manually by using the and keys on the myograph unit The speed and behaviour of the motor can be adjusted in the Setup menu by entering the code 1235 in the option line See p 25 The Mounting menu has the following functions Info Press the Info key and the display shows the F F1 F2 F3 F4 key options
6. Temp probe Output reading Analogue output Digital output Voltage Ambient temp gt 60um Auto Dual chambers Acid resistant stainless steel Max 10ml Yes Supplied with connections for gassing 100 200mN 0 01mN Automatical linear motor operated Manual Built in Ambient temp 50 C 0 1 C External Force MN Up to four output 1 0V full scale for all acquired signals user de fined Serial interface RS232 RS485 100 to 240 VAC auto 50 60Hz 15 30 C DMT Auto dual wire myograph system Model 510A rot SRC T eS Ck OS DMT A S Skejbyparken 152 DK 8200 Aarhus N Denmark Tel 45 87 41 11 00 Fax 45 87 41 11 01 www dmt dk support dmt dk sales dmt dk DMT Asia Everwin Gardens Rm 502 Block B 521 Wanping Nan Lu Shanghai 200030 China Tel 86 0 21 64869685 Fax 86 0 21 64280591 www dmt asia com sales dmt asia com support dmt asia com DMT USA Inc 1201 Peachtree Street 400 Colony Square Suite 200 630 Atlanta GA 30361 USA Tel 1 770 612 8014 Fax 1 678 302 7013 www dmt usa com support dmt usa com sales dmt usa com
7. calibration bridge balance and weight randomly on the myograph unit allowing it to be warmed up together with the myograph unit Turn on the heating in the Main menu on the Myo Interface 3 After approximately 20 30 minutes the whole system will have reached the target temperature normally 37 C Place the warmed calibration bridge and balance on the myograph unit as illustrated in fig 4 3 User manual 510A Chapter 4 29 30 4 Make sure that the tip of the transducer arm on the balance is placed in the gap between the wire and the jaw as illustrated in fig 4 4 Carefully move the calibration bridge until the tip of the transducer arm is placed freely in the gap which means it does not touch either the wire or the jaw Note The weight should not be placed on the balance yet Figure 4 3 Weight calibration setup Figure 4 4 Illustration of how to fit the balance between the wire and the gap in the support DMT Auto dual wire myograph system Model 510A Go to the Setup menu on the Myo Interface and choose to calibrate one of the transducers to change between NEAR and FAR refer to Chapter 3 Press F4 to start calibration Make sure that absolutely no force is applied on the force transducer by check ing that the tip of the transducer arm is not touching either the wire or jaw Also check that the relative force reading in the display is stable Press F3 to proceed with calibrating Carefully place th
8. for serial connection to PC i USB OKT wii NE Analogue recorder output Connection to external data acquisition system optional OROROORO Numeric keys Enter data or setting values Enter key Confirm entered numeric values or change between available choices ei ESC key Delete incorrectly entered numeric values In some submenus used to return to main menu A e ON OFF switch E nrie e Power connector MYOGRAPH Connection port for myograph unit Connection port for pH Meter optional lai e 12 V DC Port 12 DMT Auto dual wire myograph system Model 510A 1 3 Auto dual wire myograph unit Myograph chamber separator e Tu e Window at the bottom of the i myograph chamber for imaging Port for connection to Myo Allen screws for fine alignment Interface I ci of the myograph jaws gt 3 Arrow keys for manual adjustment of Motorised micropositioner the myograph jaw position S Myograph jaw connected to Ee E e Myograph jaw connected to micropositioner force transducer 1 4 Auto dual wire myograph unit with chamber cover 3 KEE e KEE Pipes for oxygen supply Suction pipes for connection to aa e Access hole for temperature probe VACUUM PUMP Orestes iii i User manual 510A Chapter 1 13 Chapter 2 Setting up 2 1 The complete myograph 510A system PC data acquisition and analysis Power supply PowerLab data acquisition system softwar
9. in residential installations This equipment generates uses and can radiate radio frequency energy and if not installed and used in accordance with the instructions may cause harmful interference to radio communications However there is no guarantee that interference will not occur in a particular installation If this equipment does cause harmful interference to radio or television reception which can be determined by monitoring the interference while turning the equipment off and on the user is encouraged to correct the interference by one or more of the following measures e Reorient or relocate the receiving antenna e Increase the separation between the equipment and receiver e Connect the equipment into an outlet on a circuit different to that which the receiver is connected to e Consult the dealer or an experienced radio TV technician for help Approvals Complies with the EMC standards EMC 89 336 EEC EN 61326 2 6 2005 EN 61000 3 2 Certified with the safety standards Directive 2006 95 EC EN 61010 1 2001 EN 61010 1 Corr 1 2003 EN 61010 1 Corr 1 2003 EN 61010 2 101 2003 DMT Auto dual wire myograph system Model 510A Certificate of Conformity DMT A S Skejbyparken 152 8200 Aarhus N Denmark hereby declares its responsibility that the following product Auto Dual Wire Myograph System Model 510A version 3 3 is covered by this certificate and marked with CE label conforms with the following standards E
10. it around under fixing screw so the wire is wound clockwise tightening the screw will then tighten the wire This procedure should result in the wire being clamped between the jaws and with near end of wire pointing towards operator fig 5 6B C e Fill the myograph chamber with PSS at room tem perature Figure 5 6 A B and C Mounting step 1 User manual 510A Chapter 5 AL Mounting step two e Using forceps to hold the handle segment transfer excised vessel from Petri dish to myograph chamber Hold the vessel as close to the proximal end as pos sible and try to mount the vessel onto the wire If the lumen is shut try one of the following possibili ties e Use the wire to gently push the lumen open blood streaming out is a good sign e Hold excised vessel about 3 mm from the cut end with one set of forceps and use the other forceps to squeeze the blood remaining in lumen out through the cut end e Pull the proximal end of the excised vessel segment along the wire such that the vessel segment acts as its own feeder to be feed into the wire into the ves sel fig AC Be careful not to stretch the vessel segment if the end of the wire catches the vessel wall C Figure 5 7 A Band C Mounting step 2 Mounting step three e Once the vessel segment is threaded onto the wire catch the free end of the wire nearest you with the forceps and move the jaws apart e While controlling the movement of the wi
11. the particular eyepiece reticule and the used magnification For such purpose a special devise called a stage micrometer is needed A stage microm eter is simply a microscope glass slide having a scale engraved on the surface A typi cal micrometer scale is 2 00 mm long engraved with divisions of 0 01 mm equalling 10um per division However micrometer glass slides with less fine divisions are also useful for calibrating a stereomicroscope to be used with the wire myograph Ocular calibration procedure 1 Decide which microscope magnification is to be used for the segment length measurements Use the largest possible fixed magnification where the eyepiece reticule scale still covers the whole gap of the myograph jaws 2 Place the stage micrometer on the microscope stage and focus on it Fit one of the division lines on the stage micrometer to one of the division lines of the reticule scale very precisely While keeping the stage micrometer absolutely fixed on the microscope stage find another position on both scales where the division lines also fit precisely Read the position of the two fit points on both scales and fill in the values in the Ocular calibration sheet Repeat the procedure twice User manual 510A Appendix 7 61 62 Ocular Calibration Sheet Dates Operatori e Microscope type and Ocular type Magnification Front lens Stage micrometer Reticular scale Position 1 Position 2 Length Positi
12. to an un derestimation of the sensitivity to acetylcholine Therefore it is recommended to apply a concentration of noradrenaline inducing 60 70 of maximal contraction response In practice this concentration is found by performing a noradrenaline concentration response curve as described in the previous section The vessel segment is exposed to the noradrenaline concentration and when the response has stabilised increasing concentrations of acetylcholine are added to relax the vessel Each concentration is applied until a steady response has been reached and then the next concentration is applied When the vessel segment is either fully relaxed or does not relax more upon increasing the acetylcholine concentration the experiment is ended User manual 510A Chapter 5 49 Protocol Prepare the following stock solutions e Acetylcholine 10 10 107M e Noradrenaline 107M 1 Mount and normalise the vessels as described in Chapter 5 2 Perform a standard start and check the vessel segment for endothelium func tion as described at the start of this chapter 3 Add noradrenaline to obtain a response around 60 of maximum determined from the previous noradrenaline concentration response curve When the con tractile response is stable add increasing concentrations of acetylcholine to the chamber using Table 6 2 as a guideline Wait for a stable contractile response or a standard time such as two minutes between each application
13. which equals 0 5 mm Each divi son on the thimble scale is 10 um If the thimble scale falls between two lines then a number between 0 and 10 um must be approximated Example 1 1 Note that the thimble has stopped at a point beyond 10 on the sleeve indicat ing 10000 um 10 mm 2 Note that there is no mark completely visible between the 10 mm mark and the thimble 3 Read the value on the thimble corresponding to the intersection with the hori zontal line on the sleeve A Reading on sleeve 10000 um B No additional mark visible O um ladini C Thimble reading 380 um tutti Total reading 10380 um SS See Example 2 Figure A7 2 Example 1 reading 10380 um 1 Note that the thimble has stopped at a point be yond 16 on the sleeve indicating 16000 um 16 mm 2 Note that this time a mark is visible between the 16 mm mark and the thimble indication 500 um 3 Read the value on the thimble corresponding to the intersection with the horizontal line on the sleeve A Reading on sleeve 16000 um B One additional mark visible 500 um C Thimble reading 280 um Figure A7 3 Example 2 Total reading 16780 um reading 16780 um User manual 510A Appendix 8 63 64 Appendix 9 System specifications Tissue size Chamber Chamber material Chamber volume Chamber gassing Chamber cover Force range Force resolution Micropositioners Weight calibration Heating Temp range Temp resolution
14. x 32 mm DMT recommends that both fuses in the fuse block are changed at the same time as it can be difficult to determine which fuse is blown To replace the fuses Use a small screwdriver to open the voltage selector block Remove the red fuse block Remove the existing fuses Insert the new fuses Replace the fuse block back into the voltage selector block Note ensure that the correct voltage for your country is displayed 2 PY Voltage selector block 58 DMT Auto dual wire myograph system Model 510A Appendix 6 Normalization theory The importance of making a Normalization before initiating an experiment with any tubular tissue segment is described in Chapter x In this appendix the mathematical rationale and calculations underlying the Normalization procedure are described in detail Mathematical calculations Let X Y be the pair of values representing the micrometer reading and force read ing respectively characterising each step in the Normalization procedure Y is the force reading at the start position of the Normalization procedure where the wires are just separated and the force reading is approximately zero Then given that ten sion on the vessel is equal to force divided by wall length the wall tension at the i th micrometer reading is calculated by pe 0 23 a a where is the microscope eyepiece reticule calibration factor in mm per division and a and a are the vessel end points wh
15. A 30361 Shanghai 200030 USA China Tel 45 87411100 Tel 86 0 21 64869685 Tel 1 770 612 8014 Fax 45 87 411101 Fax 86 0 21 64280591 Fax 1 678 302 7013 www dmt dk www dmt asia com www dmt usa com sales dmt dk sales dmt asia com sales dmt usa com support dmt dk support dmt asia com support dmt usa com User manual 510A Trademarks Introduction Until the mid 1970s most of the details about the mechanical morphological and pharmacological properties of vascular smooth muscle was obtained from studies on relatively large vessels At that time rat tail arteries were the smallest vessels to be investigated in detail due to limitations in the available in vitro techniques For example studies measuring the contraction force were routinely performed with only one of the mounting wires secured Furthermore relatively large wires 100 200 um were used which precluded the use of small vessels and the vessel segment had to be directly manipulated with the dissecting equipment causing inevitable mechanical trauma Investigations of smaller vessels were therefore limited to in vivo perfusion experiments and histological examinations In 1976 Professor M J Mulvany and Professor W Halpern described for the first time a new technique that made it possible to investigate highly isometric responses from vessels with internal diameters as small as 100 um The mounting procedure was refined twofold both ends of each mounting wire were sec
16. H meter Plastic mounting jaws DMT Item PJ 345610 With built in platinum electrodes for electrophysiological experiments e 25 um Tungsten wire DMT Item TW 345610 For mounting of ring preparations with internal diameter down to 60 um Electronic vacuum valve DMT Item VV 9937 e Standard PC system DMT Item 80150 e DMT CS 200 4 Channel Current stimulator DMT Item CS 200 Combined pulse and train generator A4 3 Myograph 510A system spare parts This section contains a complete list of standard spare parts available for the myo graph 510A For parts not listed in this Chapter or for special parts which may need to be custom made please contact DMT for further information Force transducer DMT Item FI 61 780 e Roll of 40 um stainless steel wire DMT Item SSW 123456 e High vacuum grease and grease for linear slides DMT Item HVG 1000 e Stainless steel jaws DMT Item SSJ 345610 e Mounting support pins DMT Item MO 200 e Calibration kit DMT Item CK 6127800 e Chamber cover DMT Item CC 61700 e 40 mm funnels DMT Item F 2000 User manual 510A Appendix 4 57 Appendix 5 Fuse replacement The main fuse of the myograph system is placed inside the power inlet on the Myo In terface If the fuse blows it is easily changed using the following procedure When a fuse blows and needs to be changed it is imperative that the replacement fuse is equal to the one blown The 310A system uses T1 6A 250 V 6 3
17. Myograph connection cable with a temperature probe e Power cord The shape of the AC plug varies by country be sure that the plug has the right shape for your location 2 Auto dual wire myograph unit e Calibration kit including bridge balance and 2 gram weight e Chamber cover e Chamber separator 3 Accessories e roll of 40um stainless steel wire e 1 tube of high vacuum grease e 1tubeof grease for linear slides e 4 spare screws for mounting of jaws e 2 Allen keys e 1 small screwdriver 4 Manuals e User manual for Auto dual wire myograph system Model 510A e Procedures for investigation of small vessels using small vessel myograph by Professor M J Mulvany Department of Pharmacology Aarhus University Denmark e 1CD Dissection and mounting of small vessels in wire myographs User Manual 510A Unpacking the myograph system 11 Chapter 1 System overview 1 1 Myo Interface front fanel Up and down arrow keys Scroll up and down in the menus If you are at the top Of a e menu use f to change to the previous main menu Myo Interface display e F Keys Have different but specific func tions dependent of the active main menu INFO key Push this key in any menu to display all specific F key functions i 1 2 Myo Interface rear panel 115 230 V 50 60 Hz automatic voltage selector po cupo D REGULATOR RS232 12V VC RS 232 port
18. N 61010 1 2001 Safety requirements for electrical equipment for EN61010 1 Corr 1 2003 measurement control and laboratory use EN 61010 1 Corr 1 2003 Part 1 General requirements EN 61010 2 101 2003 Safety requirements for electrical equipment for measurement control and laboratory use Part 2 101 Particular requirements for in vitro diagnostic IVD medical equipment EN 61326 2 6 2005 Electrical equipment for measurement control and laboratory use EMC requirements Part 2 6 Particular requirements In vitro diagnostic IVD medical equipment With reference to regulations in the following direc tives 2006 95 EC 89 336 EEC User manual 510A Certificate of Conformity Contents TEC NAS ee 3 EVE OG d Le TE 4 TEE 5 EMC EMI 6 KEE 6 Certificate of ContormilV isiiazi iii 7 About this CH UE 10 Unpacking the myograph SyStem c ccssccssscnsccssensennsenseensensensenssensenssenssensens 11 Chapter 1 System overview ccssccsseesscnseensenseensenssensenssenseonsenssensenssensennsenss 12 1 1 Myo Interface front fahel M nre 12 12 Myo eae rear Ee 12 1 3 Auto dual Kee e Ee AT EE 13 1 4 Auto dual wire myograph unit with chamber COVEr iii t3 Chapter 2 Setting up E 14 2 1 The complete myograph 510A System cseccececcssssereeseccsssseseecncossesasencecesseetsencecesseateeteceas 14 2 2 SOS Up Sie ry EEN 14 2 5 Tne irst WEE COMDF ANON E 15 Chapter 3 The Myo I
19. Oxygen supply Connect the small pipes on the myograph chamber cover to an adjustable oxy gen supply using small rubber tubes internal diameter 1 5 mm 4 Suction connection Connect the large pipes on the myograph chamber cover to a vacuum pump via a suction bottle and the vacuum valve as illustrated in fig 2 1 The internal diam eter of rubber tubes for connection to the pipes on the chamber cover is 2 mm An electronic vacuum valve is optional 5 pH electrode optional Connect the pH electrode to the pH port on the backside of the Myo Interface and install the software flash update as described in the purchased procedure Perform a pH calibration as described in Section 3 1 8 2 3 The first weight calibration Prior to the shipment of the 510A System it has gone through two days of continu ous testing including a final weight calibration However in order to ensure that the myograph is working at highest performance DMT recommends that a new weight calibration is performed before starting to use the myograph system The weight cali bration procedure is described in detail in Chapter 4 User manual 510A Chapter 2 15 Chapter 3 The Myo Interface 3 1 Myo Interface manual When the 510A Myo Interface is switched on the display shows that the system is starting up and initializing after which the start MYO INTERFACE _ f MODEL 510A up message depicted to the right is shown VERSION 3 30 28 02 2008 T
20. Rec 4 Rec 4 lo Rec 4 hi Change menu Start timer Stop Pause Reset Change menu Calibrate NEAR CALIBRATE NEAR FORCE F3 Apply no force Push F3 when stable Force 1715 CALIBRATE NEAR FORCE Place 2 gram on pan Push F3 when stable Force 1880 CALIBRATE NEAR FORCE DONE Push Esc Force zero 17157 Force gain 168 9 User manual 510A Chapter 3 17 18 3 2 Menus and submenus General navigation The following controls are used to display the various menus choose varying menu options and change values F1 Change to the next menu F2 F4 Have varying functions depending on the cur rent active menu Info Push this key in any menu to display all specific associated F key functions Numeric keys Enter data or setting values Enter Confirm entered numeric values or change between available choices Esc Delete incorrectly entered numeric values In some submenus used to return to main menu A Y Scroll up and down through the display as only four lines are capable of being displayed at a time Being in the top line of a submenu use A to change to the previous main menu The active line in the menu is indicated by a gt symbol Turn on message Turning on the 510A Myo Interface a message appears as shown to the right Use the and keys to adjust the display contrast setting Press F1 for proceeding to the main menu Main menu The main menu displays NEAR NEAR myograph force reading i
21. at the center of grav ity resulting in a force F_ acting on the force transducer The following two equations describe the forces working in the weight calibration system Te hy ole e Mist 8 SING 2 oS EE where r is the length of the pan arm F is the force acting on the pan arm when applying the weight F is equal acceleration of gravity times the mass of the weight r is the length of the transducer arm and F is the force acting on the force trans ducer The net torque acting at center of gravity is constant for the weight calibration setup which makes equation 1 and 2 equal making it possible to calculate the force acting on the force transducer Por olde oa IM cig 8 SING FL Ton Micia 8 sino ES SING As the length of the pan arm is 2cm the length of the transducer arm is 4 cm the weight is 2g both angles are 90 and the acceleration of gravity is 9 81 ms the force acting on the force transducer is F_ 2cm 2 gram 9 81 ms sin 90 4cm sin 90 F_ 9 81 gram m s As 1N is equal 1kg m s F is equal to F 9 81 mN 4 2 2 Weight calibration procedure This section contains a complete step by step description of how to weight calibrate the force transducers 1 Fill the myograph chamber with double distilled water and move the jaws apart Mount a 40 um stainless steel wire on the jaw connected to the force transduc ers 2 Place the
22. auses vasodilatation As the purpose is to determine the contraction sensitivity to noradrenaline the vasodilatory effect of noradrenaline is eliminated throughout the experiment by the constant presence of the B adrenoceptor antagonist propranolol Rat mesenteric arteries are densely innervated by sympathetic nerves which have a highly efficient reuptake mechanism that removes noradrenaline from the neuromuscular junction The reuptake mechanism will create a concentration gradient between the solution around the vessel segment and the receptors on the smooth muscle To correctly determine the sensitivity to noradrenaline it is necessary to eliminate this concentration gradient by performing the experiment in the presence of cocaine to block the noradrenaline reuptake To determine the sensitivity to noradrenaline the vessel segment is exposed to increasing concentrations of noradrenaline Each concentration is applied until a steady response has been reached and then the next concentration is applied When the vessel segment is fully contracted or does not response more upon increasing the noradrenaline concentration the experiment is ended Protocol Prepare the following stock solutions e Noradrenaline 10 10 107M e Propranolol 10 M e Cocaine 10 M 1 Mount and normalise the vessels as described in Chapter 5 2 Perform a standard start as described at start of this chapter 3 Incubate the vessel segment in 1 uM propranolo
23. cts damaged due to improper or inadequate packaging when returned for RMA purposes are not granted warranty coverage User manual 510A Appendix 1 53 54 Appendix 2 Service check A myograph working at optimal performance is extremely important for success when studying small blood vessels or other small tubular tissues To make sure that our customers always are dealing with first class myographs DMT offers a Myograph Service Check at a very favourable price The Myograph Service Check includes a complete separation for inspection of all mechanical and electronic parts The myograph is then reassembled adjusted and finally all electronic and mechanical parts are tested Please note that the service does not include replacement of transducers or any other needed spare parts Please contact DMT for information about prices DMT Auto dual wire myograph system Model 510A Appendix 3 Shipping instructions If the myograph system needs to be sending back for service or repair please read the following shipping instructions very carefully Before you start packing the myograph system please remember that you are dealing with very delicate equipment and therefore care must be taken DMT recommends that each part of the myograph system be wrapped individually i e with bubble wrap and placed together in a large box preferable the box you once received the myo graph system in Place the wrapped items in the middle of the box a
24. e optional Myo Interface front panel optional BED mi BEEBEDEBEBDOB Myo Interface rear panel Connection cable with temp probe Connection to oxygen supply Vaccum pump optional a 0 aa Myograph unit optional Power supply Electronic valve optional Suction bottle PRETE optional Figure 2 1 The complete Auto dual wire myograph system model 510A 2 2 Setting up step by step The chapter contains a complete step by step description of how to set up a complete myograph 510A system as illustrated in fig 2 1 1 Myograph unit Myo Interface connection Connect the myograph 510A unit to the Myo Interface using the grey 25 pin con nection cable The end of the cable with the temperature probe is used with the myograph unit 2 Myo Interface PC connection Data acquisition is possible either by connecting the Myo Interface directly to a PC or through a PowerLab data acquisition and analysis system optional I Direct PC connection Connect the Myo Interface to one of the COM ports on the PC using a serial cable not supplied with the system 14 DMT Auto dual wire myograph system Model 510A I PowerLab optional Connect the Myo Interface to the PowerLab unit using BNC cables Connect Rec 1 on the Myo Interface to Input 1 on the PowerLab Rec 2 to Input 2 etc Connect the PowerLab unit to one of the USB ports on the PC using the USB cable delivered with the PowerLab system 3
25. e 2 g weight on the pan as illustrated in fig 4 4 The force ap plied on the force transducer should mimic the stretch created by the contrac tion of a mounted ring preparation Wait until the relative force reading is stable Press F3 to finish the calibration Press Esc and go to the Main menu on the Myo Interface The force reading on the Myo Interface is now weight calibrated to an output of 9 81 mN Note If the force reading is unstable or gt 0 1 mN different from 9 81 mN repeat the weight calibration If the instability continues refer to section 4 3 9 Carefully remove the weight from the pan and repeat from step 3 to calibrate the other transducer After calibrating both transducers carefully remove the weight balance and cali bration bridge and store in the box provided The myograph system is now ready for use User manual 510A Chapter 4 31 32 4 3 Checking or replacing force transducer The myograph force transducer is a strain gauge connected in a Wheatstone bridge The force transducers are placed in separate compartment The separate compart ment provide some mechanical protection for the force transducer but the transduc ers are still very vulnerable to applied forces exceeding 1 newton 100 g or fluid run ning into the transducer compartment due to insufficient greasing of the transducer pinhole This section describes how easy it is to check the force transducer and how to replace it if necessary 4
26. e slope buffer solution and turn on stirring When the rela tive pH output in the bottom line is stable press F3 to proceed The calibration is now finished The parameters are stored in the internal memory of the Myo Interface Press ESC to return to the Setup menu Special motor speed submenu To open the special Motor Speed submenu enter the code 1235 in the option line in the Setup menu No 1235 Min Speed Initial speed Motor speed Duration The time the motor will run in Min Speed Ga before it accelerates to Med Speed Duration Med speed Med Speed Medium speed SC Duration The time the motor will run in Med Speed before it accelerates to Max Speed Max Speed Maximum speed The menu enables adjustment of the motor behaviour when using the arrow keys on the myograph unit Each setting can be adjusted between 1 and 10 where 1 is the lowest speed and 10 is the fastest speed Press F1 to return to the Setup menu Default motor speed settings are shown in the figure to the right User manual 510A Chapter 3 25 26 Chapter 4 The Auto dual wire myograph unit Chapter 4 contains a complete explanation of how to adjust calibrate and maintain the myograph 510A unit to ensure the equipment is always performing to the highest standard 4 1 Adjustment of supports A successful mounting of any kind of tubular tissue segment in the wire myograph is to a high extent dependent on perfectly ma
27. e sure that all surfaces are clean User manual 510A Chapter 4 37 Chapter 5 Getting started Chapter 5 contains some practical protocols and procedures describing how to quickly get started using the Single Wire Myograph System The chapter mostly ad dresses users who are not yet familiar with the wire myograph technique However it may contain some hints that will also be useful for the experienced myograph user 5 1 Dissection protocol for small mesenteric arteries The Single Wire Myograph technique is versatile in that a large variety of physiological and pharmacological studies of ring preparations from different species are possible The wire myograph is predominantly used for investigation of small blood vessels and as an example this chapter describes the dissection of rat mesenteric arteries 1 A laboratory rat is euthanized in accordance to the local national law and regula tions A midline laparotomy is performed to expose the mesenteric bed 2 Use scissors to remove about 10 cm of intestine together with its feeding vascu lature including part of the superior mesenteric artery Be careful not to damage the vasculature during this procedure The proximal end of the intestine section must be about 10 cm from pylorus Make a cut in the proximal end of the intes tine for later identification 3 Place the excised intestine section in a Petri dish about 9 cm in diameter coated with a 5 mm thick layer of Sylgard at the b
28. ee physiological saline solution Ca free PSS To make 1 L of Ca free PSS Use the recipe for regular PSS but omit the CaCl and add EGIA to buffer the residual Ca in solution The concentration of EGTA in the PSS should be 1 5 mM to ensure sufficient buffering 52 DMT Auto dual wire myograph system Model 510A Appendix 1 Terms of warranty Warranty DMT A S warrants to the original user that myograph systems manufactured by DMT A S will be free from defects in materials and workmanship for a period of three years after the date of delivery DMT A S will repair or replace any defective part subject to the conditions limitations and exclusions Exclusions Force and pressure transducers separately or part of myograph systems manufac tured by DMT A S are disclaimed from any warranty Limitations This warranty shall not apply to equipment subjected to accidental damage improper use alteration or deterioration Warranty on third party products will be as determined by their respective manufac turer DMT A S shall not be liable for consequential incidental special or other direct or indirect damages resulting from economic loss or property damage sustained by you or any end user from the use of the products sold or services rendered hereunder Warranty Returns A Return Material Authorisation RMA number is required for all returns This number should be clearly indicated on all returned myograph systems Produ
29. en measuring the length of the mounted vessel segment The internal circumference of the mounted vessel at the i th reading is calculated by CSIC GE A where IC is the internal circumference of the mounted vessel when the wires are just separated and is given by IC 2 7 a where d is the wire diameter For 40 um wires IC 205 6 um Using the Laplace relation the effective pressure P is calculated for each pair of readings The effective pressure is an estimate of the internal pressure which is nec essary to extend the vessel to the measured internal circumference T Bes The stepwise distension is continued until the calculated effective pressure exceeds the target transmural pressure The target value needs to be optimised for the indi vidual tissue preparation optimal active force as determined by the length tension relationship for that tissue For rat mesenteric arteries the target transmural pres sure is normally 100 mmHg 13 3 kPa IC T 100 mmHg 27 100 mmHg User manual 510A Appendix 6 59 60 An exponential curve is fitted to the internal circumference pressure data as illus trated in fig x in Chapter x Now the isobar corresponding to 100 mmHg is used to calculate the IC value from the point of interception between the function of the exponential curve and the function of the 100 mmHg isobar The normalised internal circumference IC is calculated by multiplying the internal circumfere
30. endothelium cells and subsequent relaxation of the vascular smoth muscle cells If the endothelium is undamaged by the dissection and mounting procedures then a substantial relaxation will occur With complete removal or damaged endothelium a partial relaxation or no relaxation to acetylcholine is observed It is important to note that the amount of NO or EDRF in a vessel is often dependent upon its size In certain vessels endothelium derived hyperpolarising factor EDHF can contribute more or less than EDRF and in other vessels the same stimulation with ACh can promote release of endothelium derived contracting factor EDCF Therefore it is important to check the existing literature in order to determine the expected response in your particular vessel with the given concentration of agonist Step 1 PSS 3 uM NA Stimulate for 3 minutes Step 2 Add 10 pl 5 x 10 M ACh Wait 2 minutes Step 3 Wash out 4 x with PSS Ready for experiment User manual 510A Chapter 5 47 5 6 In Vitro Experiment 1 Noradrenaline contractile response The purpose of the present protocol is to determine the sensitivity of rat mesenteric small arteries to the vasoconstrictor noradrenaline norepinephrine with a cumulative concentration response curve Background Noradrenaline norepinephrine causes contraction of mesenteric small arteries through activation of adrenoceptors whereas noradrenaline activation of B adrenoceptors c
31. es under the black covers for grease at least once a week In case of insufficient lubrication grease the slides with the original enclosed Grease for linear slides at the places marked by the arrows in fig 4 7 Kat OAI 3 OF Figure 4 7 Greasing points on the linear slides User manual 510A Chapter 4 35 4 4 4 Myograph cleaning DMT strongly recommends that the myograph chamber and surroundings should be cleaned after each experiment After a normal experiment use the following procedure to clean the myograph cham ber and supports 1 Fill the myograph chamber to the edge with an 8 acetic acid solution and allow it to work for a few minutes to dissolve calcium deposits and other salt build up Use a swab stick to mechanically clean all chamber surfaces 2 Remove the acetic acid and wash the myograph chamber and supports several times with double distilled water 3 If any kind of hydrophobic reagent have been used which might be difficult to remove using step 1 and 2 then try incubating the chamber and supports with 96 ethanol or a weak detergent solution e g Treepol 4 To remove more resistant or toxic chemicals incubate the myograph chamber and supports with 1M HCI for up to 1 hour In exceptional cases incubate the chamber and supports with an up to 3M HNO solution for about 15 minutes 5 Wash the myograph chamber and supports several times with double distilled water Important Note
32. et i P transmural pressure typically 13 3 0 00 kPa 100 mmHg IC is the normalized 0 00 internal circumference calculated from D IC o by multiplying it with a factor less d than 1 usually 0 9 Internal circumference when the wires are just touching IC 2 1 wire diameter IC 40 um wire 205 7 um Delta is the eyepiece calibration in mm div Enter the 2 delta value Microscope eyepiece reading from the far end of the mounted segment Microscope eyepiece reading from the near end of the mounted segment Microscope eyepiece reading from the far end of the mounted Microscope eyepiece reading from the near end of the mounted segment Regression coefficient for fit of X Y to an exponential curve Internal diameter of the mounted segment corresponding to a certain target transmural pressure typically 13 3 kPa 100 mmHg The micropositioner setting required to stretch the mounted segment to its normalized internal circumference Regression coefficient for fit of X Y to an exponential curve Internal diameter of the mounted segment corresponding to a certain target transmural pressure typically 13 3 kPa 100 mmHg The micropositioner setting required to stretch the mounted segment to its normalized internal circumference User manual 510A Chapter 3 21 Use the f and keys to see all values as the display is only capable of showing four lines at the time To change settings and values in
33. he display automatically shows the Main menu after several sec onds Otherwise press F1 to proceed to the Main menu When the start up message is active the A and V keys can be used to adjust the display contrast setting MAIN MENU Near 0 15 mN Far 0 02 mN Mounting Normalization Menu Act temp 36 5 C Near pos Set temp 37 0 C Far pos Heatis ON Release F Nor force IC JC IC 1 100 Info Change menu Release Together Reset motor F1 Change menu F2 Valve ON F3 Zero force F2 F4 Condensed menu Near release F2 Activate Valve F3 Yes F1 No F4 F1 Zero NEAR force F2 Zero FAR force F3 Zero both F4 Far Release F3 dia Near together Info Change menu Normalise NEAR Far Together j Normalise FAR F4 Near Reset F2 NEAR Normalization Far Reset Step 1 Time 60s F035 P 1645 7 Pressure 0 45kPa Near Seel Far 0 02 mN Act 36 8 Set 37 0 Heat ON F3 FAR Normalization Step 1 Time 60s Fonz P 1041 8 Pressure 0 28kPa Fig 3 1 Menu overview the green panels indicate the menu ap pearance options when the optional pH meter is installed DMT Auto dual wire myograph system Model 510A F1 RECORDER MENU SETUP Rec 1 NEAR Calibrate NEAR Rec l lo Valve dly 5 Rec 1 hi Contrast 55 Rec 2 Baudrate 9600 Rec 2 lo Option Rec 2 hi Rec 3 Rec 3 lo Rec 3 hi
34. his chapter Repeat 1 x Stimulus 1 amp 2 Wash out KPSS 10 uM NA 4 x with PSS Stimulate for 3 minutes Wait 5 minutes Stimulus 3 Wash out PSS 10 uM NA 4 x with PSS Stimulate for 3 minutes Wait 5 minutes Stimulus 4 Wash out KPSS 4 x with PSS Stimulate for 3 minutes Wait 5 minutes Stimulus 5 Wash out KPSS 10 uM NA 4 x with PSS Stimulate for 3 minutes Ready for experiment DMT Auto dual wire myograph system Model 510A 5 5 Endothelium function The reasons for checking endothelium function may include 1 To check whether the relaxing function of the endothelium is intact The procedure is performed to make sure that the endothelium is not damaged during the dissec tion or mounting procedure 2 If an experiment requires removal of the endothelium this procedure is useful to check whether the endothelial cells were successfully removed The procedure can be performed after the vessel segment has been heated equili brated and normalised Preferably the procedure should be done after performing a standard start to make sure that the vessel segment is viable The present procedure is for use with rat mesenteric arteries Another procedure may be needed for other animal species and tissue or vessel types Principles of checking endothelium function Stimulating a vessel segment with acetylcholine causes a release of nitric oxide NO also known as EDRF from the
35. iment remove the chamber cover from the myograph and turn on the vacuum pump and vacuum valve for about 10 seconds Wait to turn off the oxygen supply until turning off the vacuum pump Wipe off any buffer remaining on the outside of the pipes using a piece of paper 4 4 2 Force transducer The force transducer is the most delicate and fragile component of the myograph system Therefore careful handling is necessary One of the jaws in each myograph is connected to the transducer pin The transducer pin enters the myograph chamber through a pinhole in the chamber wall located be low the surface level of the buffer as illustrated in fig 4 6 To prevent the buffer from running into the transducer house the hole is filled with high vacuum grease As a part of daily maintenance it is very important to inspect the greasing of the transducer hole before starting any experiment Insufficient greasing causes damage and mal function of the force transducer Important e DMT recommends use of the high vacuum grease once a week to seal up the transducer hole by frequently use e DMT takes no responsibilities for the use of any other kinds of high vacuum grease than the one to be purchased from DMT e DMT takes no responsibilities for any kind of damage applied to the force trans ducer Figure 4 6 Transducer pin hole to be sealed up with high vacuum grease DMT Auto dual wire myograph system Model 510A 4 4 3 Linear slides Check the linear slid
36. in the Mounting menu Press Info again to return to the Mounting menu or one of the F keys to proceed with one of these options Press F1 to proceed to the Normalization menu Press F2 to open the Release menu Press F1 to release NEAR myograph jaws Press F4 to release FAR myograph jaws The jaws will then move apart until the distance between the jaws is equal to the release set value Press F3 to open the Together menu Press F1 to move NEAR myograph jaws together Press F4 to move FAR myograph jaws together Press F4 to open the Reset menu Press F1 to reset the NEAR myograph motor Press F4 to reset the FAR myograph motor DMT recommends that both motors are reset before any experiment is performed DMT Auto Auto dual wire myograph system Model 510A Info Change menu Release Together Reset motor F2 F1 Near release F2 F3 F4 Far Release F3 Near together Far Together F4 Near Reset Far Reset Normalization menu The Normalization menu displays Norm pres Norm time N Nor force F Nor force IC 1C 00 Target transmural pressure usually 13 3 kPa 100 mmHg Time setting in seconds defining the duration of each step in the automatic TRO et normalization Norm me 60s N Nor fi 2 0 mN NEAR myograph force reading in mN COS FAR myograph force reading in mN i oe IC o is the internal circumference SE corresponding to a certain targ
37. ing any direct contact between the scissor and artery fig 5 4 A B Figure 5 4 Removal of vein Figure 5 5 A B and C Cutting free the artery to be studied Ata DMT Auto dual wire myograph system Model 510A 9 Clean the artery by removing any remaining adipose or connective tissue Gently pull away adipose or connective tissue to make the connective tissue membrane become visible Cut the membrane to remove the tissue 10 Cut the distal end of the artery section to be investigated Afterwards cut the proximal end while ensuring that the vessel segment has the correct length fig 5 5A C 5 2 Mounting protocol for small arteries The procedure involves attaching the mounting wires to jaws which are in turn mount ed on the force transducer This force transducer is capable of measuring with a sensitivity of about 0 01 mN 1 mg but can be damaged if the applied force exceeds about 1 N 100 g Therefore care must be taken to avoid pressing the jaws too hard together A movement of 20 um after they have touched is sufficient to hold the wires clamped Mounting step one e Cut lengths of 40 um wire 2 2 cm long Mount one wire on left hand jaw of myograph as follows e Holding wire at far end place centre of wire between Jaws and screw jaws together so that the wire is clamped fig 5 6A Note do not close the jaws too hard against each other See above Bend the far end of the wire towards the left and wrap
38. ins procedures describing general as well as daily maintenance of the myograph unit e g adjustment of Supports weight calibration of the force trans ducer and cleaning instructions Chapter 5 describes how to get started using the wire myograph system This in cludes a complete dissection and mounting procedure for small mesenteric arteries and a few basic pharmacological experiments Appendices contain additional information about Normalization theory ocular cali bration myograph service shipping instructions system specifications equipment lists accessories and spare parts and fuse replacement DMT Auto dual wire myograph system Model 510A Unpacking the myograph system Please take a few minutes to carefully inspect your new 510A wire myograph system for damage which may have occurred during handling and shipping If you suspect any kind of damage please contact DMT immediately and the matter will be pursued soon as possible If the packing material appears damaged please retain it until a possible claim has been settled We recommend that you store the packing material for any possible future transport of the Wire myograph system In case of transport and the original packing material is unavailable please contact DMT Sales Department for advice and packing instructions After unpacking your new 510A wire myograph system please use the following list to check that the system is complete 1 Myo Interface unit e
39. l lowing characteristics fig 5 3 38 DMT Auto dual wire myograph system Model 510A Second Branch Figure 5 1 Branching of the mesenteric arteries Figure 5 2 Removal of adipose tissue around the area of interest Figure 5 3 Distinguishing between artery and vein User manual 510A Chapter 5 39 40 e The branch points of arteries are V shaped whereas those of veins are more U shaped e The arterial wall contains a thick layer of smooth muscle cells compared to the vein wall which only contains a single or a few layers of smooth muscle cells The histological difference is clearly visible in the stereomicroscope e If you still have difficulty and the vein and artery still contain some blood then try to move the blood forward by very gently squeezing the vessels with a for ceps In the artery the blood will run back quickly whereas in the vein the blood will run back very slowly if it even does so Note it is important that you per form this on vessels other than those you will use as this procedure damages the vessels 8 Dissect away the vein using scissors to cut the adipose and connective tissue between the artery and vein One method is to cut the vein in one position and af terwards gently to pull the vein away from the artery In this way a fine membrane of connective tissue becomes visible between the adipose tissue and the artery Carefully cut the fine membrane to remove the vein and adipose tissue while avoid
40. l add 5 uL of 10 Mto 5 mL PSS in chamber and 3 uM cocaine add 15 uL of 10 M to 5 mL PSS in chamber for at least 10 minutes 4 Add increasing concentrations of noradrenaline into the bath use Table 6 1 as a guideline Wait for a stable contractile response or a standard time such as 2 minutes between each application DMT Auto dual wire myograph system Model 510A D Lue0N D of sas ss E In calculating the NA in the myograph chamber the ee volume of no radrenaline is ignored 5 7 In Vitro Experiment 2 Acetylcholine relaxation curve The purpose of the present protocol is to determine the sensitivity of the endothelium dependent vasodilator acetylcholine in noradrenaline pre contracted rat mesenteric small arteries Background Acetylcholine causes relaxation of rat mesenteric small arteries by activating of mus carinic M3 receptors at the endothelial cell layer leading to release of endothelium derived relaxing factors Rat mesenteric arteries do not show spontaneous tone in the wire myograph which is why it is necessary to first induce a contraction to be able to observe the relaxa tion to acetylcholine In this protocol the contraction is induced by noradrenaline The required concentration of noradrenaline needs to be optimised since a too low concentration makes it impossible to evaluate the relaxation On the other hand it may be difficult to relax super maximally contracted arteries which may lead
41. lades one being wider than the other A grounding type plug has two blades and a third round grounding pin The wide blade or the third prong is provided for your safety If the provided plug does not fit into your outlet consult an electrician for replacement of the obsolete outlet e Be advised that different operating voltages require the use of different types of line cord and attachment plugs Check the voltage in your area and use the correct type See the table below Line plug according to standard 110 125 V UL817 and CSA C22 2 No 42 220 230 V CEE 7 page VII SR section 107 2 D1 IEC 83 page C4 240 V BS 1363 of 1984 Specification for 13A fused plugs and switched and unswitched socket outlets Protect the power cord from being walked on or pinched particularly at power plugs and the point where they connect to the apparatus Refer all servicing to qualified service personnel Servicing is required when the apparatus has been damaged in any way such as the power supply cord or plug is damaged liquid has spilled onto or objects have fallen into the apparatus the apparatus has been exposed to rain or moisture does not operate normally or has been dropped User manual 510A Safety EMC EMI This equipment has been tested and found to comply with the limits for a Class B Digital device pursuant to part 15 of the FCC rules These limits are designed to provide reasonable protection against harmful interference
42. n mN FAR FAR myograph force reading in mN Act Temp Actual temperature probe reading in C Set Temp Heating temperature setting in C Heat is Actual heating status on off Use the f and keys to see all values as the display is only capable of showing four lines at the time Temperature setting To change the temperature setting move the Set Temp line to the top of the display Use the numeric keys to enter a new temperature setting and press Enter to confirm DMT Auto dual wire myograph system Model 510A Myo Interface Model 510A Version 3 30 OP ALO OS Main Menu ISMN 82 mmigd Act Temp 26 5 C Set Temp 37 0 C Heat is on Heating control To turn on heating move the Heat is line to the top of the display and use Enter to switch between on and off The Main Menu has the following functions Info F1 F2 F3 F4 Press the Info key and the display shows the F key options in the Main menu Press Info again to return to the Main menu or one of the F keys to proceed with one of these options Press F1 to proceed to the Recorder menu Press F2 to open the Valve activation Menu An electronic vacuum valve is available as optional to the standard manual valve The electronic vacuum valve is connected to the regulator port on the backside of the myo graph Press F1 to activate the electronic vacuum valve Press F4 to close the electronic vacuum valve
43. nce corresponding to 100 mmHg IC po by a factor k The factor is for rat mesenteric arteries 0 9 Again this value should be optimised for the particular tis sue preparation being used by a length tension curve IC k IC The normalised internal lumen diameter is then calculated by fe 1 TU The micrometer reading X at which the internal circumference of the normalised ves sel is set to is calculated by IC IC X X 2 DMT Auto dual wire myograph system Model 510A Appendix 7 Calibration of eyepiece reticule Principles of ocular calibration The purpose of calibrating the eyepiece reticule is to determine a conversion factor 6 allowing the microscope to be used for measuring vessel seg ment lengths mounted in the wire myograph 6070 0 90 10 O dm D Ap So Several types of eyepiece reticules are available for such a purpose The most simple and yet very useful type is a horizontal scale as illustrated in fig A7 1 Figure A7 1 Horizontal eyepiece reticule scale The basic principle is to use the eyepiece reticule typically consisting of 50 100 divi sions to measure the length of an object in terms of reticular divisions spanned by the object Having the conversion factor specific for the eyepiece reticule and used magnification the length of the object in mm is easily calculated All reticules need to be calibrated in order to determine the specific conversion factor characteristic for
44. nd fill out the Surroundings with chips of expanded polystyrene Important Before closing the box make sure that no enclosed items can be shaken around as transport by road or air from time to time can be quite roughly Address the box to DMT A S Skejbyparken 152 DK 8200 Aarhus N Denmark Make sure that all four sides of the box are marked fragile or similar Make an indi cation on the top of the box that it contains goods returned for repair service Customers outside the EC must further enclose a pro forma invoice stating that the box contains goods being returned for repair or service If arranging transportation through a courier please keep in mind the high value of the myograph system and that a standard insurance provided by the courier in most cases is insufficient to cover damage or loss of the myograph system In most cases an additional insurance coverage is needed User manual 510A Appendix 3 55 Appendix 4 Myograph accessories and spare parts This section contains a complete register of equipment needed to set up a Single Wire Myograph System In addition the chapter contains a list of special myograph 310A accessories and spare parts Besides the main focus on development and manufacturing DMT has specialised in offering our costumers first class laboratory equipment needed for a Single Wire Myograph setup at very competitive prices Please contact DMT Sales Department for further product information a
45. nd prices A4 1 General myograph equipment This sedtion contains a complete checklist of laboratory equipment needed when set ting up a Single Wire Myograph system Dissection stereo microscope including ocular micrometer and stage micrometer DMT recommends the Zeiss Stemi 2000 Stereo Microscope e Mounting forceps DMT Item DF 3000 DMT recommends Dumont Medical No 5 tip 0 10 mm x 0 06 mm Dissection scissors DMT Item DS 1000 DMT recommends Geuder G 19745 8 cm straight trabeculum e Pipettes DMT recommends CappAerou pipettes s Light source DMT recommends Schott Cold Light Source either Model KL 200 or Model KL 1500 LCD e Water bath including heater DMT recommends Julabo 5 I open bath circulator with plexiglass bath tank for internal temperature applications e Glass bottle 2 L Vacuum pump DMT recommends a membrane vacuum pump having a volume of at least 6 litres per minute Dissection petri dish DMT Item PD 2000 DMT recommends a 9 cm glass Petri dish coated with a 5 mm Sylgard polymer layer DMT Auto dual wire myograph system Model 510A A4 2 Myograph 510A system accessories This section contains a list of special accessories available for the Auto dual wire myograph system Model 510A ADI PowerLab data acquisition system Including Chart data acquisition and analysis software e DMT Normalization module DMT Item MLSO65 Chart add on software e pH Meter Including pH electrode and p
46. ne ani 54 Appendix 3 Shipping INStrucCtiONs ccsccsscenscnsecnsenssensecssensenssenssensenssensens 55 Appendix 4 Myograph accessories and spare parts 11 11121020000 56 A4 1 General myograph COUIDIN aero 56 A4 2 Myograph 510A system aCCessorles NENNEN 57 A4 3 Myograph 510A system spare parts on Di Appendix 5 Fuse replacement 1 11 111xsvs0r0sare rara KEREN KEREN KENE KEN 58 Appendix 6 Normalization theory see KEREN KENNEN KEN KEN KEN KEREN KEE kk 59 Appendix 7 Calibration of eyepiece retiCule c cssscsssessseenseenseenseeneeees 61 Appendix 8 How to read a millimetre micrometer 211 21x 21x gt 63 Appendix 9 System specifications 1 211 211 2r0s arrese irene iena 64 User manual 510A Contents 10 About this manual This manual contains a complete list of procedures describing how to install maintain and get started using the Auto dual wire myograph system model 510A version ie Pro Chapter 1 provides an overview of the construction and basic features of the Myo In terface and the Auto dual wire myograph unit Chapter 2 describes step by step how to set up a complete 510A wire myograph sys tem including accessories Chapter 3 is a complete manual to the Myo Interface The chapter describes in detail the construction of the menu system and how to use all the features of the Auto dual wire myograph system Chapter 4 conta
47. nterface cc ccssscesscenseesseenseenseenseenseecssenseennsesseensenees 16 EN e Geht e E 16 3 2 MENUS and SUDMECNUS iaia 18 Chapter A The Auto dual wire myograph UNIt ccsccssccnscnseensenssnseensensennes 26 A LAdiUSIMEnNt Or SUPPO EE 26 4 2 Calibration of force dee e EE 28 eee D diete ler en gel e sooi 28 4 2 2 Weight calibration e reel 29 43 Checking OF replacing Torce en EE 2 4 3 1 Simple transducer Check ri 32 43 2 FORCE transducer feplaGeMent a tranne 32 4 4 Myograph maintenance ssessssonsnnrnnnnnrnrrsennnnnnnnnnnrrrnrnnnnnnnnnnnrennnnnnnnnnnnneneennnnnnnnnnnnnnennnnn nnmnnn 34 44 1 My graph chamber e TE 34 4 4 2 Force transducer rune 34 Fea PMC A TE 39 4 4 4 Mersin mnene 36 Chapter 5 Getting started 1 11 2s02srs0rerariane ENKEN einen 38 5 1 Dissection protocol for small mesenteric ALtEVieS ccccesesessseevsseseveeceeueesueseesaeueesaeueesaeeeenas 38 5 2 Mounting protocol for small aFeleS i rri iaia 41 De IN OMIT CIE VON EE 45 e 200 NANNE EEEE AEEA EARTE ENANS ES NANEN EENE AKRE E 46 So OG NING EITC EE 4T 5 6 In Vitro Experiment 1 Noradrenaline contractile response 48 5 7 In Vitro Experiment 2 Acetylcholine relaxation CUFVE rire 49 DMT Auto dual wire myograph system Model 510A Appendix 1 Terms of warranty 11 211021020r0 errare ener irene 53 Appendix 2 Service check 11 211 1022svrsss creare aaa ne ie
48. nual 510A Chapter 5 51 25x Concentrated PSS To make 1 L concentrated PSS Solution 1 Chemical MW g mol Conc mmol L Conc g L NaCl 58 44 118 99 173 85 KCI 14 56 4 69 8 7 5 CaCl 2H O 147 02 2 50 9 20 Solution 2 Chemical MW g mol Conc mmol L Conc g L MgSO HO 246 48 ES 1 23 KH2PO 136 09 1 18 4 02 Solution 3 Chemical MW g mol Conc mmol L Conc g L EDTA 372 24 0 03 0 25 1 Dissolve the chemicals for solution 1 in about 800 mL double distilled H O in a 1000 mL graduated bottle Dissolve the chemicals for solutions 2 and 3 in 75 mL double distilled H O in individually cylinders Gently heat solution 3 to dissolve the EDTA 2 Solution 2 and 3 is added to solution 1 and the graduated bottle is filled with additional double distilled H O to reach a final volume of 1000 mL Before use 3 Dilute the 25 x PSS stock solution 1 25 with double distilled HO 4 Add 1 091 g L Glucose 2 100 g L NaHCO 5 Aerate the solution with carbogen 95 O 5 CO for at least 20 minutes If necessary wait further for the pH of the buffer to reach pH 7 4 High Potassium physiological saline solution KPSS To make 1 L of KPSS Use the recipe for regular PSS but replace the desired concentration of NaCl with KCI For example to make 60 mM KPSS Solution 1 Chemical MW g mol Conc mmol L Conc g L NaCl 58 44 64 86 3 179 KCI 174 56 58 82 4 39 MgSO HO 246 48 1 17 0 29 KH PO 136 09 LAS 0 16 Calcium fr
49. o the standard manual vacuum valve Valve delay is the time in seconds that the valve stays open when activated Contrast The actual contrast setting in the display Option The option line allows access to submenus using special codes To change any of the settings in the Setup menu move the line to be changed to the top line of the display e Press Enter to change between calibration of NEAR myograph force FAR myograph force or pH e Use the numeric keys to change the valve delay or dis play contrast settings and press Enter to confirm e Enter the specific access code and press Enter to go to the submenu Timer Menu Hour 10 Minute O Second 0 Info Change menu Start timer Stop Pause Reset Setup Menu Calibrate Valve dly Contrast Option User manual 510A Chapter 3 23 24 The Setup menu has the following functions Info Press the Info key and the display shows the F key options in the Setup menu Press Info again to return to the Setup menu or one of the F keys to proceed with one of these options F1 Press F1 to proceed to the Main menu F4 Transducer calibration Having chosen which one of the myograph force transducers to calibrate NEAR or FAR press F4 to enter the window shown to the right Make sure that no force is applied on the force transducer and when the relative reading at the bottom line of the window appears stable press F3 to proceed Caref
50. ographs and organ tissue baths Driven by our global customer base our number one goal is to develop and manufacture first class research equipment within the fields of physiology and pharmacology DMT Auto dual wire myograph system Model 510A Safety The Auto dual wire myograph system has been designed for use only in teaching and research applications It is not intended for clinical or critical life care use and should never be used for these purposes nor for the prevention diagnosis curing treatment or alleviation of disease injury or handicap e Do not open the unit the internal electronics pose a risk of electric shock e Do not use this apparatus near water e To reduce the risk of fire or electric shock do not expose this apparatus to rain or moisture Objects filled with liquids should not be placed on the apparatus e Do not block any ventilation openings Install in accordance with the manufacturer s instructions e Do not install near any heat sources such as radiators heat registers stoves or other apparatus that produce heat e Only use attachments and accessories specified by the manufacturer e Unplug this apparatus during lightning storms or when unused for long periods of time e This apparatus must be grounded e Use a three wire grounding type cord similar to the one supplied with the product e Do not defeat the safety purpose of the polarized or grounding type plug A polarized plug has two flat b
51. on1 Position 2 Length 1 div div mm div 2 div div mm div 3 div div mm div Calculations 1 Calculate the length between the two positions on the stage micrometer by sub tracting the value of position 1 from the value of position 2 Dualply the length in divisions with the length of each division to get the length in mm 2 Calculate the length between the two positions on the reticule scale by subtracting the value of position 1 from the value of position 2 3 Calculate the mean length value of both the stage micrometer and the reticular scale 4 Calculate the conversion factor 5 Mean Stage Micrometer Length mm Mean Reticule Scale div DMT Auto dual wire myograph system Model 510A Appendix 8 How to read a millimetre micrometer Sleeve scale Thimble scale ba Figure A7 1 Overview of the micrometer parts actual reading 20000 um 20 mm Sleeve scale The micrometer sleeve scale has a total length of 25 mm divided into fifty equal parts Each part of division above the horizontal line represents 1 mm where each fifth line is marked by a longer line and a number which designates the length in mm Each part of division below the horizontal line is placed between each 1 mm mark scale above the horizontal line and represents 0 5 mm Thimble scale The thimble is divided into fifty equal parts and one complete rotation of the thimble is indicated by the smallest division on the sleeve
52. ond the jaws as even a small extension will af fect the Normalization procedure In case of excess of vessel on the far side of the jaws then move the jaws together again and remove excessive tissue using a forceps as described in mounting step four A better method for the skilled operator is to move the jaws slightly apart and use scissors to make a small slit in the vessel wall where the vessel is clamped B Figure 5 12 A and B Mounting step 7 DMT Auto dual wire myograph system Model 510A 5 3 Normalization The importance of normalizing the preparation is three fold 1 Experiments with elastic preparations like vessels can only have meaning if they are performed under conditions where the size is clearly defined 2 Clearly defined conditions are required in pharmacological experiments as the sensitivity of preparations to agonists and antagonists is dependent on the amount of stretch 3 The active response of a preparation is dependent on the extent of stretch which makes it important to set the preparation to an internal circumference giving maximal response The aim of the normalization procedure is to stretch the segment to a so called normalized internal circumference IC defined as a set fraction of the internal cir cumference IC that a fully relaxed segment would have at a specified transmural pressure For small rat arteries the target transmural pressure is typically LOO mmHg 13 3 kPa Principles of
53. ottom to hold the fixing pins Immedi ately fill the Petri dish with cold PSS which has been well aerated with carbogen The dissection is performed without further oxygenation of the PSS The cold tem perature ensures the artery is relaxed during the mounting procedure 4 Pin down the proximal end of the intestine section on the left hand side of the Petri dish without stretching the vessels Pin down the remaining of the intestine section in an anti clockwise direction In this configuration proximal end at the left side distal end at the right side and running anti clockwise from proximal to distal side the feeding vasculature is on the far side of the intestine and the veins are usually uppermost 5 Select the vessel segment to be investigated fig 5 1 First time myograph users are recommended to start dissecting and mounting vessel segments from the first or second branch from the superior mesenteric artery internal diameter 200 300 um 6 Use high quality forceps and ocular dissection scissors to dissect the vessel segment of interest Start cutting through the mesenteric membrane along both sides of the vessel about 1 2 mm from the vessel To avoid accidentally cutting the artery always cut along the length of the vessels and never perpendicular to them fig 5 2A B T Dissect away as much adipose tissue as needed around the vessels to distin guish between the artery and vein The artery can easily be identified by the fo
54. perties of the vessel segment 2 Check that responses to different types of stimuli are normal in appearance and thereby ensuring that the functionality of the vessel segment has not been dam aged during the dissection or mounting procedures 3 Ensure that the tension development gives an effective active pressure that is above the chosen accepted value usually 13 3 kPa 100 mmHg The standard start is performed after the vessel segment has been heated equili brated and normalised The present procedure is suitable for rat mesenteric arter ies Another procedure may be needed for other animal species and tissue or vessel types Principles of the standard start procedure The standard start procedure consists of a series of five stimuli and washout periods The first two stimuli are performed using a mixture of KPSS and 10 uM noradrena line to give a maximum contractile response The third stimulus is performed using a mixture of PSS and 10 uM noradrenaline to give a maximum pure agonist mediated a adrenoceptor contraction The fourth stimulus is performed using KPSS to give a depolarising contractile response this stimulus also includes a component from neurally released noradrenaline The final stimulus is performed using a mixture of PSS and 10 uM noradrenaline All solutions are preheated to 37 C and aerated with a mixture of 95 O and 5 CO before use Instructions for making the necessary solutions are described at the end of t
55. re with the forceps use the other forceps to gently pull the ves sel segment along the wire until the area of interest is situated in the gap between the jaws The near end of the vessel segment shall lie about 0 1 mm inside the jaw gap to insure no point of contact fig 5 8A e Still controlling the free wire end with the forceps move the jaws together to clamp the wire and in one movement secure the wire under the near fixing screw on the left hand jaw Again in a clock wise direction so that tightening the screw also tightens the wire fig 5 8B 42 DMT Auto dual wire myograph system Model 510A B Figure 5 8 A and B Mounting step 3 Mounting step four e Using forceps gently rub the vessel segment on the far side of the jaw to separate any excess ves sel segment from the area of interest clamped in the gap between the jaws fig 5 9A Make sure that the vessel segment is separated as close as possible to the jaws The excessive vessel segment is finally dissected free and removed from the myo graph chamber fig 5 9B C C Figure 5 9 A B and C Mounting step 4 Mounting step five e Move the jaws apart fig 5 10A Take a second wire holding it about one third down from the far end using a forceps Align the wire parallel with the vessel segment such that the wire can be passed into the far end of the lumen Gently feed the wire through the lumen of the vessel segment in one movement using
56. s Be very careful using step 3 and 4 repeatedly times as strong reagents may cause extreme damage to the myograph unit e After cleaning ALWAYS check that the greasing around the transducer pin is suf ficient to keep the buffer solution away from the transducer compartment In cases of red or brown discolorations appearing on the chamber sides or on the supports the following cleaning procedure will work in most cases 1 Incubate the myograph chamber and supports for 30 minutes with 2Oul of a 2mM T 1210 Tetrakis 2 pyridylmethyl ethylenediamine solution dissolved in double distilled water 2 Use a swab stick to mechanically clean all the affected surfaces during the last 15 minutes of the incubation period 3 Wash the myograph chamber and supports several times with double distilled water 4 Incubate the myograph chamber with 96 ethanol for 10 minutes while continu ing the mechanical cleaning with a swab stick 5 Remove the ethanol solution and wash a few times with double distilled water Incubate the myograph chamber and supports with an 8 acetic acid solution for 10 minutes and continue the mechanical cleaning with a swab stick 36 DMT Auto dual wire myograph system Model 510A 6 Wash the myograph chamber and supports several times with double distilled water Important Notes e In exceptional cases it may be necessary to demount the supports for cleaning Supports and the myograph chamber individually to mak
57. tching supports The supports are matched prior to the shipment but daily use of the myograph system and greasing of the transducer pinhole will over time create a need for an adjustment of the supports Adjustment of the supports is performed using the following step by step procedure Please note that the amount of force on the screws should be very little to avoid breaking the threads The procedure is illustrated in fig 4 1 1 Carefully loosen screw D on the top of the support connected to the force transducer Align the horizontal support and carefully tighten the screw again 2 Loosen screw D on the top of the support connected to the linear slide Align the horizontal support matching the force transducer connected support as care fully as possible and gently tighten the screw again 3 Loosen screw C on the linear slide to roughly match the linear slide support to the force transducer support in the horizontal plane as illustrated in fig 4 1 1 Tighten the screw before proceeding with step 4 4 The plate B on which the linear slide is mounted is balanced on top of a small stainless steel ball making it possible to finely adjust the linear slide support in all vertical and horizontal planes using the four Allen screws A Use the four Allen screws to make the final horizontal fig 4 1 1 and vertical fig 4 1 2 adjustments to match the linear slide support to the force transducer support The correct matching of
58. the Normalization menu move the line to be changed to the top of the display Use the numeric keys to enter a new setting value and press Enter to confirm The Normalization menu has the following functions Info Press the Info key and the display shows the Sg F key options in the Normalization menu Change menu Normalise NEAR Normalise FAR Press Info again to return to the Normalization menu or one of the F keys to proceed with one of these options F1 Press F1 to proceed to the Recorder menu F2 F3 After entering all necessary settings and values in the Normalization menu press NEAR Normalization Step 1 Time 60s F085 P 1645 7 Pressure 0 45kPa F2 to start normalization of the segment in the NEAR myograph F3 FAR Normalization F3 to start Normalization of the segment in IRE the FAR myograph F 0 14 P 1041 8 Pressure 0 28kPa Press ESC to cancel the normalization As the Normalization reaches the target transmural pressure usually 13 3 kPa 100 mmHg the Normalization position is calculated and the micropositioner is turned automatically to the correct position The results of the Normalization are shown afterwards in the Normalization menu Recorder menu The 510A Myo Interface has four analog output ports at Recorder Menu the front panel for connection to a data recording system a Aa Each of these output ports can be programmed to repre Rec dhi 20 sent either NEAR m
59. the first mounted wire as a guide fig 5 10B C Hold the wire at a point at least 10 mm from the vessel to prevent the vessel being stretched during the manoeuvre Be careful not to touch the lumen of the vessel with the end of the wire and when pushing the wire end through the near end of the lumen Once the wire has success fully passed through the lumen of the vessel seg ment place the wire in a position which ensures sufficient length for the wire to be secured both at the near and far fixing screws on the right hand De jaw Figure 5 10 A Band C Mounting step 5 User manual 510A Chapter 5 Mounting step six e Carefully move the jaws together while ensuring that the second mounted wire lies underneath the first one secured on the left hand jaw fig 5 11A The proce dure clamps the second wire to prevent it from damag ing the vessel segment when securing the wire to the right hand jaw connected to the transducer Secure the near end of the wire in a clockwise direction under the far fixing screw on the right hand jaw fig 5 11B Figure 5 11 AandB Mounting step 6 Mounting step seven e Secure the far end of the wire under the near fixing screw on the right hand jaw Again the wire is passed clockwise around the screw stretching the wire as the screw is tightened fig 5 12A B Move the jaws apart to slightly stretch the vessel segment Make sure that the vessel on the far side of the jaws does not extend bey
60. the normalization procedure In practice the normalisation is performed by distending the segment stepwise and measuring sets of micrometer and force readings fig 5 13 step 1 4 These data are converted into values of internal circumference um and wall tension T mN mm respectively Plotting wall tension against internal circumference reveals an exponential curve and by applying the isobar curve corresponding to 100 mmHg IC is calculated from the point of intersection using the Laplace relation fig 5 14 IC is calculated from IC oo by multiplying a factor giving an internal circumference at which the active force production as well as the sensitivity to agonists of the segment is maximal For rat mesenteric arteries the factor is 0 9 but both this factor as well as the transmural pressure has to be optimised for each particular segment The normalized internal diameter is calculated by dividing IC with x Appendix 6 contains a complete description of the mathematical rationale and cal culations of the nNormalization procedure Force mNimm 100 200 300 200 500 600 700 800 900 Internal Circumference pom Time min Figure 5 13 Illustration of the stepwise Figure 5 14 Illustration of the expo normalization procedure nential curve fitting and determina tion of IC User manual 510A Chapter 5 45 5 4 Standard start The purpose of performing a standard start is to 1 Re activate the mechanical and functional pro
61. the supports is illustrated in fig 4 1 3 and 4 Important Be careful not only to continuously tighten the Allen screws when doing the final adjustments Always remember to loosen the Allen screw placed diagonal to the Allen screw being tightened or else there is a high risk of damaging the myograph frame DMT Auto dual wire myograph system Model 510A Figure 4 1 Illustration of how to adjust supports User manual 510A Chapter 4 4 2 Calibration of force transducer As a part of the general maintenance of the myograph DMT recommends that the myograph is weight calibrated at least once every month DMT also recommends that the myograph is weight calibrated every time the system has been moved or has not been used for a long period of time 4 2 1 Principles of weight calibration Weight calibrating the force transducer is based on simple physics the net torque acting on a balance when applying a certain amount of weight The magnitude of the torque T about a point of rotation P is defined by Sh T r F sing where r is the distance from the point of rotation to the point on the object where the force F is acting with the angle of 0 Applying the physics to the weight calibration setup is illustrated in fig 4 2 Transducer Arm TA Figure 4 2 Thereotical principle of the weight calibration 28 DMT Auto dual wire myograph system Model 510A Applying the weight on the pan arm creates a net torque acting
62. ully place the 2 gram weight on the load pan and when the relative reading at the bottom line of the window appears stable press F3 to proceed The calibration is now finished The readings in the two bottom lines are the calibration parameters stored in the internal memory of the Myo Interface Press ESC to return to the Setup menu For further information about the calibration proce dure see chapter 4 DH calibration optional Having chosen to calibrate the pH settings press F4 to enter the window shown to the right The first line shows the value of the offset buffer first buffer solution which is always 7 00 The second line shows the temperature of the buffer solution The temperature is an important parameter in the calibration formula and is ob tained automatically by placing the myograph temperature probe in the buffer solution The third line shows the output from the pH probe as raw data from the A D converter Place the pH meter electrode and temperature probe in the offset buffer solution and turn on stirring When the relative pH output in the bottom line is stable press F3 to proceed DMT Auto dual wire myograph system Model 510A Info Change menu Calibrate Near Calibrate NEAR FAR Force Apply no Force Push F3 when stable Force 1938 In the first line is now shown the value of the slope buffer second buffer solution which is always 4 00 Place the pH and temperature probes in th
63. ured under tension without the need to manipulate the vessel segment Segments of small vessels could now be atraumatically mounted as ring preparations in a myograph for recording of highly isometric force measurements During the late 1970s some improvements were made to the myograph and in 1981 a new dual myograph that allowed simultaneous testing of two vessels was introduced In parallel the technique became more acknowledged worldwide resulting in a growing interest in the myograph systems In 1986 the growing demand resulted in the foundation of the private company J P Trading with the purpose of making the myograph systems commercially available worldwide At the same time J P Trading initiated a comprehensive improvement programme for the existing myograph systems as well as a development programme of new myograph systems in close co operation with Professor M J Mulvany and the University of Aarhus During the late 1980s and through the 1990s several improvements were applied to the myograph systems such as a new mechanical design a more robust transducer and new electronic systems In addition new systems were introduced like the Automatic Dual Myograph 510A the Dual Myograph 410A the Multi Myograph 610M and the Confocal Myograph 120CW In 2000 J P Trading changed its company structure and became known as DMT Today DMT is one of the world s leading designers and manufacturers of wire myographs pressure myographs culture my
64. yograph FAR myograph temperature e or pH meter The default settings are E Ee REC 3110 gee Rec 3 hi 2 BO Rec 4 Rec 4 lo Rec 4 hi NEAR myograph force output FAR myograph force output Temperature output The full scale output on the recorder is one volt In the Recorder menu it is possible to change the value that cor responds to O V lo and 1 V hi DMT Auto dual wire myograph system Model 510A Timer menu The Timer menu is a feature making it possible to set an alarm based on a count down from a predefined set value of hours minutes and or seconds To set the count down time move the line to be programmed hour minute or second to the top of the display Use the numeric keys to enter the time value and press Enter to confirm The Timer menu has the following functions e Info Press the Info key and the display shows the F key options in the Timer menu e Press Info again to return to the Timer menu or one of the F keys to proceed with one of these options F1 Press F1 to proceed to the Setup menu F2 Press F2 to start the count down after the time values have been entered in the Timer menu F3 Press F3 to stop or pause the count down F4 Press F4 to reset entered time values or to turn off the alarm Setup menu The Setup menu displays Calibrate Choose which sensor to be calibrated NEAR FAR myograph force or pH Valve dly An electronic vacuum valve is available optional t

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